Department of Biology, College of Natural Sciences, Kyungpook National University, Daegu 702-701, Republic of Korea.
Acta Histochem. 2013 Jun;115(5):475-80. doi: 10.1016/j.acthis.2012.11.003. Epub 2012 Dec 3.
Although several matrix metalloproteinases (MMPs) have been implicated in testis development, the presence of MMP-13 protein has not been directly substantiated in the male avian gonads. In this study, we examined the expression patterns of MMP-13 and MMP inhibitors, TIMP-1 and TIMP-2, in immature (4weeks), pre-pubertal (16weeks), and mature (1year) chicken testes. Using RT-PCR analysis, we observed that MMP-13 mRNA was expressed in immature testis. In Western blot analysis, the expression level of MMP-13 protein peaked in the immature testes during marked tissue remodeling, whereas it gradually decreased during testis maturation. High expression levels of TIMP-1 (34-kDa) and TIMP-2 (55-kDa) were detected only in immature and pre-pubertal testes and not in adult testis. Four different forms of TIMP-2 protein were differentially detected in the testes of growing and adult chicken. Using immunohistochemistry we localized both secreted and intracellular forms of MMP-13, TIMP-1, and TIMP-2 proteins. These proteins were temporally and spatially distributed in growing and adult testes, and all their expression levels were similar to the expression profile of Western blot results. These findings suggest that age-related changes of MMP-13 with balance of TIMPs act in concert to effect the controlled testicular remodeling and maturation.
虽然几种基质金属蛋白酶(MMPs)已被牵涉到睾丸发育中,但 MMP-13 蛋白在雄性禽类性腺中的存在尚未得到直接证实。在这项研究中,我们研究了 MMP-13 和 MMP 抑制剂,TIMP-1 和 TIMP-2,在未成熟(4 周)、青春期前(16 周)和成熟(1 年)鸡睾丸中的表达模式。通过 RT-PCR 分析,我们观察到 MMP-13 mRNA 在未成熟睾丸中表达。在 Western blot 分析中,MMP-13 蛋白的表达水平在组织重塑明显的未成熟睾丸中达到峰值,而在睾丸成熟过程中逐渐降低。TIMP-1(34-kDa)和 TIMP-2(55-kDa)的高表达水平仅在未成熟和青春期前的睾丸中检测到,而在成年睾丸中未检测到。在生长和成年鸡的睾丸中,检测到四种不同形式的 TIMP-2 蛋白。通过免疫组织化学,我们定位了 MMP-13、TIMP-1 和 TIMP-2 蛋白的分泌型和细胞内型。这些蛋白在生长和成年睾丸中具有时间和空间分布,它们的所有表达水平都与 Western blot 结果的表达谱相似。这些发现表明,MMP-13 与 TIMPs 的平衡随年龄变化,共同作用于控制睾丸重塑和成熟。